Fertilization and development of mouse oocytes injected with isolated sperm heads
Fertilization and development of mouse oocytes injected with isolated sperm heads
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DOI：
10.1095/biolreprod55.4.789
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发表时间：
1996-10-01
影响因子：
3.6
通讯作者：
Yanagimachi, R
中科院分区：
文献类型：
--
作者：
Kuretake, S;Kimura, Y;Yanagimachi, R
为了确定精子在显微外科注射到卵母细胞中进行正常受精之前是否必须结构完整，将完整的精子以及通过超声处理与尾部分离的精子头部单独注射到卵母细胞中。当整个精子固定后立即注射时，大多数卵母细胞受精并正常发育。无论有或没有 Triton X-100，超声处理都能斩首超过 95% 的精子。尽管所有断头精子都通过活/死精子染色被诊断为“死亡”，但分离的精子头（细胞核）在注射到卵母细胞中时可以参与正常的胚胎发育。分离的精子头（细胞核）参与正常胚胎发育的能力在不适合质膜完整精子存活的冷冻保存条件下得以维持。这些结果表明：1) 精子不需要在胞质内注射时结构完整，2) 质膜和顶体膜以及所有尾部成分对于正常胚胎发育不是必需的，至少在小鼠中是这样，3) 维持精子核遗传完整性所需的冷冻保存条件不如保持质膜完整所需的条件严格。 精子还活着。
To determine whether spermatozoa must be structurally intact before microsurgical injection into oocytes for normal fertilization, intact spermatozoa, as well as sperm heads separated from tails by sonication, were individually injected into oocytes. When whole spermatozoa were injected immediately after their immobilization, the majority of the oocytes were fertilized and developed normally. Sonication in the presence or absence of Triton X-100 decapitated more than 95% of spermatozoa. Although all decapitated spermatozoa were diagnosed as ''dead'' by live/dead sperm staining, separated sperm heads (nuclei) could participate in normal embryo development when injected into the oocytes. The ability of isolated sperm heads (nuclei) to participate in normal embryo development was maintained under cryopreservation conditions that were not suitable for the survival of plasma membrane-intact spermatozoa. These results indicate that 1) spermatozoa do not need to be structurally intact for intracytoplasmic injection, 2) the plasma and acrosomal membranes and all tail components are not essential for normal embryo development, at least in the mouse, and 3) the cryopreservation conditions required for maintenance of the genetic integrity of sperm nuclei are less stringent than those necessary for keeping plasma membrane-intact spermatozoa alive.